Test if antitumor T cells use a putative super-enhancer in the Il1rl2-Il1rl1 intergenic region to form "innate-like" responses to cytokines

测试抗肿瘤 T 细胞是否使用 Il1rl2-Il1rl1 基因间区域中的假定超级增强子对细胞因子形成“先天样”反应

• 批准号: 9585298
• 负责人: ADAM J ADLER
• 金额: $ 19.94万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-04 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9585298
• 关键词: 3-Dimensional; Affinity; Agonist; Antigen Receptors; Antigens; Avidity; Binding; Binding Sites; Bioinformatics; Bypass; CD8-Positive T-Lymphocytes; Cancer Patient; Cardiovascular Diseases; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Loop; Collaborations; Consensus; Cytotoxic T-Lymphocyte-Associated Protein 4; Data; Data Set; Elements; Enhancers; Epitopes; Family; Family member; Foundations; Funding; Future; Generations; Genes; Genomics; High-Throughput Nucleotide Sequencing; Immune; Immune checkpoint inhibitor; Immunosuppressive Agents; Immunotherapeutic agent; Inflammation; Intercistronic Region; Interferon Type II; Interleukin-1; Interleukin-12; Interleukin-18; Interleukin-2; Joints; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Molecular; Mus; Natural Killer Cells; PDCD1LG1 gene; Pathway interactions; Process; Publications; Publishing; RNA Polymerase II; Research; Role; Schedule; Science; Site; Stat5 protein; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Tumor Antigens; Tumor Immunity; Work; anti-PD-1; base; cancer immunotherapy; cancer therapy; cell killing; cell motility; chromatin immunoprecipitation; chromosome conformation capture; cytokine; experience; genome editing; immune function; meetings; member; neoplastic cell; novel; patient subsets; programs; promoter; receptor function; response; treatment response; tumor; tumor growth; tumor microenvironment

Project Summary The new generation of T cell-based cancer immunotherapeutics, best exemplified by the checkpoint inhibitors anti-CTLA-4 and anti-PD-1/PD-L1, can elicit therapeutic antitumor immunity in cancer patients. Nevertheless, these therapeutic responses are often not durable or only experienced in a subset of patients, likely owing to a variety of immunosuppressive mechanisms employed by tumors to subvert the action of antitumor T cells. For instance, T cells expressing antigen receptors (TCRs) that recognize MHC restricted-tumor epitopes with high avidity/affinity have a propensity to undergo functional inactivation or deletion, and hence cytolytic effector T cells (CTL) localized within tumors generally have a limited ability to recognize and be triggered by tumor epitopes. Hence, strategies to amplify weak tumor epitope recognition, or even induce TCR-independent T cell activation intratumorally, might boost T cell-mediated antitumor efficacy. In this regard, we previously found that effector T cells programmed with antitumor promoting costimulatory agonists can be triggered to degranulate and secrete the tumoricidal cytokine IFN-γ through a TCR-independent process that involves stimulation with a STAT-activating cytokine (IL-2 or IL-12) plus an IL-1 family member (IL-33 or IL-36). This “innate-like” response is akin to how innate immune cells such as NK cells can be triggered through cytokine combinations such as IL-12 plus IL-18, and may help explain why the presence of IL-36 in the tumor microenvironment limits tumor growth. Further, engaging this 2-cytokine triggering pathway will be an effective means to boost the efficacy of cancer immunotherapies. In analyzing the mechanism of this 2-cytokine pathway using costimulated CD8+ T cells treated with the IL-2 + IL-36 combination, it was found that IL-2 primes for IL-36 responsiveness by rapidly inducing IL-36R mRNA (encoded by the Il1rl2 gene) in a JAK/STAT-dependent manner. Chromatin immunoprecipitation with high throughput sequencing (ChIP-seq) revealed that IL-2 induces STAT5 binding to five separate STAT/GAS consensus elements located throughout a large ~75 kb intergenic region between Il1rl2 and Il1rl1 (encoding IL-33R that is also induced by IL-2). Strikingly, RNA polymerase II bound the same sites prior to IL-2 stimulation. These results, in concert with a previous bioinformatic analysis of publically available ChIP-seq data sets generated from IFN-γ-expressing T cells, forms the basis of the novel overarching hypothesis that will be tested in this project that the Il1rl2-Il1rl1 intergenic region contains a super-enhancer (SE) that serves as a molecular hub to facilitate cross-talk between STAT-activating and IL-1 family cytokines during “innate-like” antitumor T cell responses.

项目摘要 以T细胞为基础的新一代癌症免疫疗法，检查点抑制剂是最好的例证 抗CTLA-4和抗PD-1/PD-L1可诱导肿瘤患者产生治疗性抗肿瘤免疫。不过， 这些治疗反应通常不持久或仅在一小部分患者中体验到，可能是由于 肿瘤用来破坏抗肿瘤T细胞作用的多种免疫抑制机制。为 例如，表达抗原受体(TCR)的T细胞识别高MHC限制性肿瘤表位 亲和力/亲和力有经历功能失活或缺失的倾向，因此细胞溶解效应T 定位于肿瘤内的细胞(CTL)通常对肿瘤的识别和被肿瘤触发的能力有限 表位。因此，扩增弱肿瘤表位识别，甚至诱导TCR非依赖性T细胞的策略 瘤内激活，可能会增强T细胞介导的抗肿瘤效果。在这方面，我们之前发现 用抗肿瘤促进共刺激激动剂编程的效应器T细胞可以被触发 通过不依赖TcR的过程脱颗粒和分泌杀瘤细胞因子干扰素-γ，该过程涉及 使用激活状态的细胞因子(IL-2或IL-12)加上IL-1家族成员(IL-33或IL-36)进行刺激。这 “类先天”反应类似于NK细胞等先天免疫细胞如何通过细胞因子被触发 如IL-12和IL-18的组合，并可能有助于解释为什么IL-36在肿瘤中的存在 微环境限制了肿瘤的生长。此外，参与这种2-细胞因子的触发途径将是一种有效的 提高癌症免疫疗法疗效的手段。在分析这种2-细胞因子的作用机制时 利用共刺激途径用IL-2+IL-36联合处理CD8+T细胞，发现IL-2 通过快速诱导IL-36R mRNA(由IL1rl2基因编码)在小鼠体内启动IL-36反应 依赖于JAK/STAT的方式。染色质免疫沉淀高通量测序(CHIP-SEQ) 发现IL-2诱导STAT5与分布于全身的五个独立的STAT/GAS共识元件结合 在IL1rl2和Il1rl1之间有一个大的~75kb的基因间隔区(编码IL-33R，也由IL-2诱导)。 引人注目的是，在IL-2刺激之前，RNA聚合酶II结合了相同的位点。这些结果，与一个 先前对从表达干扰素γ的T细胞产生的公共可用的CHIP-SEQ数据集的生物信息学分析 细胞，形成了新的总体假说的基础，该假说将在本项目中进行测试，即Il1rl2-Il1rl1 基因间隔区包含一个超级增强子(SE)，作为促进串扰的分子枢纽 STAT激活和IL-1家族细胞因子在“先天类”抗肿瘤T细胞反应中的作用。